An international comparability study on quantification of total methyl cytosine content.
Various methods have been developed for quantitative analysis of DNA methylation. However, there is currently no reference analysis system regarding DNA methylation with which other analytical approaches can be compared and evaluated. A standard measurement system that includes reference methods and reference materials may improve comparability and credibility of data obtained from different analytical environments. In an effort to establish a standard system for measurement of DNA methylation, the Korea Research Institute of Standards and Science (KRISS) coordinated an international comparison study among different national metrology institutes. An initial stage of the study involved an intercomparison regarding quantitative measurement of total methyl cytosine contents in artificially constructed DNA samples. The measurement principle involved measurement of dNMP contents following enzymatic hydrolysis of DNA samples. Results of the study showed good comparability among four of five participants and close agreement with reference values assigned by the coordinating laboratory. Conflicting data from one participant may have resulted from incomplete hydrolysis of samples due to use of insufficient amounts of enzymes. These results indicate that comparable and accurate results can be obtained from different measurement environments if digestion conditions are controlled appropriately and valid calibration systems are employed.
Yang
Inchul
I
Health Metrology Group, Korea Research Institute of Standards and Science, 1 Doryong-dong, Yusung-gu, Daejon 305-340, Republic of Korea.
Kim
Sook-Kyung
SK
Burke
Daniel G
DG
Griffiths
Kate
K
Kassir
Zena
Z
Emslie
Kerry R
KR
Gao
Yunhua
Y
Wang
Jing
J
Foy
Carole A
CA
Pardos-Pardos
Ana Carmen
AC
Ellison
Steve
S
Domann
Paula Jane
PJ
Fujii
Shin-Ichiro
S
Park
Sang-Ryoul
SR
D003596:Cytosine / Q000031:analogs & derivatives* / Q000032:analysis*; D019175:DNA Methylation; D003854:Deoxyribonucleotides / Q000737:chemistry; D019075:Electrophoresis, Capillary; D006868:Hydrolysis; D012015:Reference Standards
D003854:Deoxyribonucleotides; D003596:Cytosine